
UNIT DEFINITION
One unit (U) is defined as the amount of enzyme which produces 1 μmol of PNP per min under the conditions described below.
CAS # 9001-42-7
REAGENTS PREPARATION
Reagent I: 100 mM PBS, pH 7.0.
Reagent II: 20 mM p-Nitrophenyl-α-D-glucopyranoside (PNPG).
Reagent III: 200 mM Na2CO3.
Reagent IV: 200 mM, pH 7.0 potassium
phosphate buffer, contain 0.05% Tween 20 and 1 mM EDTA.
Sample: Dilute the sample to 0.006-0.022 U/mL by Reagent IV.
PROCEDURE
- Add 1mL Reagent I, 0.5 mL Reagent II to a test tube, and preincubate for 5 min at 37 ℃.
- Add 0.5 mL sample and mix, react for 15 min in 37 ℃, then add 2 mL reagent III to stop the reaction.
- Record the ΔAs at 400 nm in a spectrophotometer.
- Exchange the adding order of reagent III and sample, and measure the blank absorbance ΔAb.
∆A = ∆As – ∆Ab

C: Enzyme concentration (mg/mL)
t: Reaction time (15 min)
18.1: Millimolar extinction coefficient of p-Nitrophenol under 400 nm (cm2/μmol)
REFERENCES
- Ranscher, E. et al (1986). Analyt. Chem.324:
- Kruse – Jarres, J.D. et al (1989). J. Clin. Chem. 27: 103
This enzyme is useful for diagnostic tests for the determination of α-amylase.
The Certificate Of Analysis (COA) & Material Safety Data Sheet (MSDS) Is A Signed Document That Includes The Storage Temperature,
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